Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Biodegradation of Endocrine Disrupting Chemicals by Genetic Transformants of Phlebia tremellosa Using Manganese Peroxidase Gene from Trametes versicolor

구름버섯 망간 과산화효소를 도입한 아교버섯 형질전환체에 의한 내분비장애 물질의 생분해

  • Kum, Hyun-Woo (Department of Biochemistry, Division of Life Sciences, Kangwon National University) ;
  • Kim, Myung-Kil (Division of Wood Chemistry and Microbiology, Korea Forest Research Institute) ;
  • Choi, Hyoung-T. (Department of Biochemistry, Division of Life Sciences, Kangwon National University)
  • 금현우 (강원대학교 생명과학부 생화학과) ;
  • 김명길 (국립 산림과학원 화학미생물과) ;
  • 최형태 (강원대학교 생명과학부 생화학과)
  • Published : 2009.03.31
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Abstract

Endocrine disrupting chemicals (EDCs) disturb animal hormonal system even at very low concentrations, and finally give harmful effects to human through the food web. A white rot fungus Phlebia tremellosa isolated in Korea, was reported to have good degrading activity against the endocrine disrupting phthalates. However, this fungus has very low manganese peroxidase (MnP) activity under various culture conditions while laccase and lignin peroxidase activities were high. We have isolated an MnP cDNA from Trametes versicolor which was involved in the degradation of EDCs, and constructed an MnP expression vector to use in the genetic transformation of P. tremellosa in order to get higher MnP producing strains. Many transformants had integrated expression vector in their chromosomal DNAs, and showed increased MnP activity. One of two transformants showed increased degradation of 4 EDCs (70${\sim}$88%) than the wild type (30${\sim}$45% degradation rates), and showed twice better removal of estrogenic activities generated by the EDCs than the wild type.

내분비장애물질은 분해가 매우 어려워 자연계에서 먹이그물을 통하여 사람에게 농축 전달된다. 이들은 정상적인 내분비계에 혼란을 일으키며, 특히 성호르몬의 작용에 많은 피해를 준다. 이를 효율적으로 분해하고 이들의 에스트로겐 활성을 제거하고자 백색부후균의 하나인 아교버섯(Phlebia tremellosa)을 활용하여 4가지 내분비계 장애물질의 분해에 대한 실험을 수행하였다. 아교버섯의 manganese peroxidase (MnP) 활성을 높이기 위하여 구름버섯의 MnP 유전자를 아교버섯에 도입하여 형질전환체를 확보하였으며 이들은 유전적으로 MnP 활성을 안정되게 나타냈다. 내분비 장애물질을 분해하는 조건에서 내분비장애 물질에 따라 30${\sim}$45%의 분해율을 보인 야생형에 비하여 이 형질전환체들 중 T5는 70${\sim}$88%의 분해율을 보였으며 에스트로겐 활성의 제거에도 약 2배 향상된 능력을 보였다.

Keywords

References

  1. 강애리, 최형태, 송홍규. 2008. Trametes versicolor에 의한 bisphenol A 생분해의 최적조건. 미생물학회지 44, 37-42
  2. 김윤정, 송홍규, 최형태. 2008. 기계충버섯 형질전환체를 이용한 비스페놀 A의 분해와 에스트로겐 활성 제거. 미생물학회지 44, 199-202
  3. 여수민, 김명길, 최형태. 2008. 아교버섯 형질전환체를 이용한 내분비장애 물질의 분해. 미생물학회지 44, 10-13
  4. Baborova, P., M. Moder, P. Baldrian, K. Cajthamlova, and T. Cajthaml. 2006. Purification of a new manganese peroxidase of the white-rot fungus Irpex lacteus, and degradation of polycyclic aromatic hydrocarbons by the exzyme. Res. Microbiol. 157, 248-253 https://doi.org/10.1016/j.resmic.2005.09.001
  5. Bhatti, H.N., N. Akram, and M. Asgher. 2008. Optimization of culture conditions for enhanced decolorization of cibacron red FN-2BL by Schizophyllum commune IBL-6. Appl. Biochem. Biotechnol. 149, 255-264 https://doi.org/10.1007/s12010-007-8123-x
  6. Cheong, S., S. Yeo, H.G. Song, and H.T. Choi. 2006. Determination of laccase gene expression during degradation of 2,4,6-trinitrotoluene and its catabolic intermediates in Trametes versicolor. Microbiol. Res. 161, 316-320 https://doi.org/10.1016/j.micres.2005.12.001
  7. Chhabra, M., S. Mishra, and T.R. Sreekrishnan. 2008. Mediatorassisted decolorization and detoxification of textile dyes/dye mixture by Cyathus bulleri laccase. Appl. Biochem. Biotechnol. 151, 587-598 https://doi.org/10.1007/s12010-008-8234-z
  8. Colborn, T., D. Dumanoski, and J. Meyers. 1996. Our stolen future. Dutton, New York, USA
  9. Hirano, T., Y. Honda, T. Watanabe, and M. Kuwahara. 2000. Degradation of bisphenol A by the lignin-degrading enzyme, manganese peroxidase, produced by the white-rot basidiomycete, Pleurotus ostreatus. Biosci. Biotechnol. Biochem. 64, 1958-1962 https://doi.org/10.1271/bbb.64.1958
  10. Hwang, S.S., H.T. Choi, and H.G. Song. 2008. Biodegradation of endocrine-disrupting phthalates by Pleurotus ostreatus. J. Microbiol. Biotechnol. 18, 767-772
  11. Leem, Y., S. Kim, I.K. Ross, and H.T. Choi. 1999. Transformation and laccase mutant isolation in Coprinus congregatus by restriction enzyme-mediated integration. FEMS Microbiol. Lett. 172, 35-40 https://doi.org/10.1111/j.1574-6968.1999.tb13446.x
  12. Palma, C., M.T. Moreira, G. Feijoo, and J.M. Lema. 1997. Enhanced catalytic properties of MnP by exogenous addition of manganese and hydrogen peroxide. Biotechnol. Lett. 19, 263-267 https://doi.org/10.1023/A:1018313825723
  13. Schonfelder, G., B. Flick, E. Mayr, C. Talsness, M. Paul, and I. Chahoud. 2002. In utero exposure to low doses of bisphenol A lead to long-term deleterious effects in the vagina. Neoplasia 4, 98-102 https://doi.org/10.1038/sj.neo.7900212
  14. Shin, E.H., H.T. Choi, and H.G. Song. 2007. Biodegradation of endocrine-disrupting bisphenol A by white rot fungus Irpex lacteus. J. Microbiol. Biotechnol. 17, 1147-1151
  15. Tsutsumi, Y., T. Haneda, and T. Nishida. 2001. Removal of estrogenic activities of bisphenol A and nonylphenol by oxidative enzymes from lignin-degrading basidiomycetes. Chemosphere 42, 271-276 https://doi.org/10.1016/S0045-6535(00)00081-3
  16. Yeo, S., M.K. Kim, and H.T. Choi. 2008. Increased expression of laccase by the addition of phthalates in Phlebia tremellosa. FEMS Microbiol. Lett. 278, 72-77 https://doi.org/10.1111/j.1574-6968.2007.00971.x
  17. Yeo, S., N. Park, H.G. Song, and H.T. Choi. 2007. Generation of a transformant showing higher manganese peroxidase (MnP) activity by overexpression of MnP gene in Trametes versicolor. J. Microbiol. 45, 213-218